• Title/Summary/Keyword: HPLC Chromatography

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Isolation and Characterization of MMP-1 Inhibitor Peptide from Crataegus pinnatifida Bunge in Fibroblast Cell Line HS68 Cells (아가위(Crataegus pinnatifida Bunge)로 부터 HS 68세포의 MMP-1에 대한 저해활성 물질의 분리)

  • Lee, Se-Young;Chun, Hyug;Cho, Hong-Yun;An, Jeung-Hee
    • Applied Biological Chemistry
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    • v.46 no.1
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    • pp.60-65
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    • 2003
  • MMP-1 inhibitory compounds were isolated from 120 Korean traditional edible plants. UP- 1 activity significantly increased linearly with increasing UVB dose in normal human foreskin fibroblast HS68 cell, showing maximum activity at approximately 35 $mJ/cm^2$, whereas in HaCaT cell, normal human keratinocyte, no increase was observed. Maximum secretion of MMP-1 after UVB treatment occurred around 36-48 k after treatment. MMP-1 inhibitory compound isolated from cold-water fraction of Cataegus pinnatifida Bunge showed the mort potent activity. The MMP-1 inhibitory compound was deduced as a peptide based on the fact that pronase digestion decreased the activity whereas periodate oxidation did not. The most potent UP- 1-inhibitory protein, CP-2Va-2, showing an activity of 88.5% against MMP-1, was isolated through sequential column chromatography on DEAE-Toyopearl 650C, Butyl-Toyopearl 650M, and Bio-Gel P-30. Molecular weight of CP-2Va-2 determined through high performance liquid chromatography and SDS PACE was 19 and 20 kDa. respectively, signifying a monomeric structure.

Studies on the Changes of Taste Compounds during Soy Paste Fermentation (III) (된장 숙성 중 정미 성분의 변화에 관한 연구(III))

  • 김미정;이혜수
    • Korean journal of food and cookery science
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    • v.9 no.4
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    • pp.261-265
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    • 1993
  • For the purpose of supplying the information to improve the acceptability of soy paste as the condi-ment, the changes of peptide were determined. The results were as follows; Average peptide length were decreased. It was 102 at 0 day, 15 at 10 day and 4.1 at 180 day. Peptide fraction were the same as in 60 day and 180 day. Low molecular weight peptide were not changed greatly during fermention. Peptide identified in 180 day fermentation were Ala-Ser, Gly-Glu, Glu-Ser, Asp-Glu, Asp- Tyr, Asp-Ala-Ser, Ala-Ser-Glu, Glu-Ser-Ala, and Ala-Lys-Met. In the characteristics of bitter peptide in 180 day fermentation, soy paste itself didn't show bitter taste', solvent extration fraction I'showed bitter taste. After gel chromatography, fraction I, fraction II and fraction III were obtained and fraction II were bitter peptide of low molecular weight. After gel chromatography', solvent extration fraction 2'(water extration) were divided into fraction IV, V, VI,VII and VIII. Fraction IV, V and VI showed bitter taste. Amino acids composition of the fractions showing bitter taste were like that; fr. 1: Glu- (Asp, Pro, Val, lie or Leu)-Met fr. II Pro-(Glu, Val, Phe)-lle or Leu fr. IV: Glu-(Asp, Ala, Tyr, Leu of lie)-Phe fr. V: Ala-(Met, Glu, Pro)-lle or Leu fr. VI: Asp-(Phe, Ser, fly)-Val.

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Multiresidue Determination of Tetracyclines in Eggs using Liquid Chromatography with Ultraviolet Detection (액체크로마토그래피를 이용한 계란 중 테트라사이클린계 항생물질의 동시분석법 개발)

  • Lee, Sang-Hee;Shim, You-Sin;Choi, Yoon-Hee;Lee, Beom-Gil;Kim, Hyun-Ju;Shin, Dong-Bin
    • Journal of Food Hygiene and Safety
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    • v.22 no.4
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    • pp.370-374
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    • 2007
  • An analytical method for the simultaneous determination of four tetracycline (oxytetracycline, tetracycline, chlortetracycline, doxycycline) in egg samples was developed and validated using liquid chromatography with ultraviolet detection. Egg samples were extracted by the liquid-liquid extraction based on acetonitrile. The chromatographic separation was achieved on a reverse phase C8 column with gradient elution using a mobile phase of 20 mM oxalic acid (pH 1.5)/acetonitrile. The procedure was validated according to the Food Drugs Administration guideline determining accuracy, precision, and limit of detection. Mean recovery of tetracyclines from spiked egg samples (50, 100, 200, 400, and $800{\mu}g/kg$) were 78.8-109.3%. Linearity in concentration range of $50-800{\mu}g/kg$ was obtained with the correlation coefficient $(r^2)$ of 0.994-0.999. The intra- and inter-day precision (relative standard deviation; RSD) was between 0.3-12.8 and 0.2-11.7%, respectively. Limit of detection (LOD) and limit of quantification (LOQ) for the investigated tetracyclines were 30 and $50{\mu}g/kg$ depending on egg samples, respectively. This method was reliable, sensitive, economical and suitable for routine monitoring of tetracycline residues in dairy egg.

Conversion of Ginseng Saponin with the Enzyme Produced by Rhizopus sp. (Part 1) Confirmation of Conversion of Ginsenoside- Rb$_1$to Ginsenoside-Rd (Rhizopus sp.가 생산하는 효소에 의한 인삼 Saponin의 전환 (제1보) Ginsenoside-Rb$_1$에서 Ginsenoside-Rd로의 전환확인)

  • 김상달;서정훈
    • Microbiology and Biotechnology Letters
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    • v.10 no.4
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    • pp.267-273
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    • 1982
  • Among 12 kinds of ginsenosides in ginseng saponin, ginsenoside-Rb$_1$was contained the most abundantly. But ginsenoside-Rd which is similar to ginsenoside-Rb$_1$in structure, was known to be superior to ginsenoside-Rb$_1$pharmaceutically. In order to convert ginsenoside-Rb$_1$into ginsenoside-Rd by microbial enzyme treatment, a Rhizopus sp. was selected among various strais of molds found in rotten ginseng roots. Enzyme was prepared from the extract of wheat bran koji culture by ammonium sulfate precipitation (1.0 sat'd) and succeeding ammonium sulfate fractionation method (0.6-0.9 sat'd). For the purpose of use as substrate, saponins were purified by the several purification steps from alcohol extract of red ginseng roots. We obtained the total saponin which was composed of 36.5% of ginsenoside Rb$_1$, 12.2% of ginsenoside-Rd and other ginsenosides. For increase of ginsenoside-Rb$_1$ component ratio, we also obtained further purified ginsenoside-Rb group saponin containing 54.5% of ginsenoside-Rb$_1$, 1.1% of ginsenoside- Rd and other ginsenosides from purified the total saponin. In the enzymatic reaction system including the total saponin or the ginsenoside-Rb group saponin, we confirmed the specific conversion of ginsenoside-Rb$_1$to ginsenoside-Rd proportionally and no change of any other ginsenoside patterns by thin layer chromatography and high performance liquid chromatography.

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Elution Behavior of Pd(II) - Isonitrosoethylacetoacetate Imine Chelates by Reversed Phase High Performance liquid Chromatography (역상 액체 크로마토그래피에 의한 Pd(II) - Isonitrosoethylacetoacetate Imine 유도체 킬레이트들의 용리 거동)

  • Kim, In-Whan;Shin, Han-Chul;Lee, Man-Ho;Yoon, Tai-Kun;Kang, Chang-Hee;Lee, Won
    • Analytical Science and Technology
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    • v.5 no.4
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    • pp.389-399
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    • 1992
  • Liquid Chromatographic behavior of Pd(II) in Isonitrosoethylacetoacetate lmine, $Pd(IEAA-NR)_2$ (R=H, $CH_3$, $C_2H_5$, $n-C_3H_7$, $C_6H_5-CH_2$, $n-C_4H_9$) chelates were investigated by reversed-phase HPLC on Micropak MCH-5 column using methanol/water as mobile phase. The optimum conditions for the separation of $Pd(IEAA-NR)_2$ chelates were examined with respect to the effect of the flow rate, sample solvent, mobile phase strength and column temperature. It wass found that metal chelates were properly eluted in an acceptable range of capacity factor value($0{\leq}log\;k^{\prime}{\leq}1$). The dependence of the logarithm of capacity factor(k') on the volume fraction of water in the binary mobile phase was examined. Also, the dependence of k' on the liquid-liquid extration distribution ratio($D_c$) in methanol-water/n-alkane extration system was investigated. Both kinds of dependence are linear, which susggests that the retention of the electroneutral metal chelate is largely due to the solvophobic effect. Standard adsorption enthalpy changes (${\Delta}H^{\circ}$) and standard adsorption entropy changes (${\Delta}S^{\circ}$) of Pd(II) Isonitrosoethylacetoacetate imine chelates on Micropak MCH-5 column were calculated by measuring capacity factor with changing temperature of the column.

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Characterization of Anti-Complementary Polysaccharides Isolated from Fruit Wine Using Korean Pears (배를 이용한 과실주로부터 분리한 항보체 활성화 다당의 특성)

  • Choi, Jung-Ho;Shin, Kwang-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.1
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    • pp.63-69
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    • 2011
  • To characterize the polysaccharides which exist as soluble forms in Korean traditional alcoholic beverages, the polysaccharides were isolated from Korean pear wine and their anti-complementary activities were examined. The main polysaccharide, PW-1 was purified to homogeneity from the crude polysaccharide (PW-0) in pear wine by size exclusion chromatography using Sephadex G-75. Molecular mass of PW-1 was estimated to be 150 kDa and it contained significant proportion of mannose (81.8%) and 5 different minor component sugars such as arabinose (1.2%), galactose (2.7%), glucose (8.5%), galacturonic acid (5.3%) and glucuronic acid (0.5%). These analyses indicated that the main polysaccharide in pear wine was mainly present as a mannan which had originated from the cell walls of fermenting yeasts. On the other hand, PW-1 showed potent anti-complementary activity in a dose-dependent fashion. Identification of C3 activation products by the crossed immunoelectrophoresis using anti-human C3 and anti-complementary activity of PW-1 in $Ca^{++}$-free condition suggested complement activations by PW-1 from Korean pear wine occur via both classical and alternative pathways.

Stability of Danggwisu-san (Dangguixu-san) Water Extract, a Herbal Medicine, Under Various Storage Conditions (보관조건에 따른 당귀수산의 성분 변화)

  • Do, Ho-Jeong;Shin, Ye-Sle;Lee, Jae-Hwan;Ahn, Yong-Jun;Ha, In-Hyuk;Lee, Yoon-Jae;Kim, Mi-Riong;Ko, Won-Il;Song, Seol-Hee;Chung, Hwa-Jin;Lee, In-Hee;Lee, Jae-Woong;Kim, Eun-Jee;Kim, Min-Jeong
    • Journal of Korean Medicine Rehabilitation
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    • v.26 no.4
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    • pp.1-8
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    • 2016
  • Objectives Danggwisu-san (Dangguixu-san) is a herbal prescription frequently used to treat pain or swelling caused by contusion. To determine the expiration period through scientific methodology, stability of Danggwisu-san (Dangguixu-san) water extract, a herbal medicine, was examined under various storage conditions and periods. Methods Danggwisu-san (Dangguixu-san) was stored either at room temperature ($23{\pm}2^{\circ}C$), under a refrigerating condition ($4^{\circ}C$) or under a freezing condition ($-18{\pm}2^{\circ}C$) for 0, 1, 2, 3 or 4 weeks and then freeze-dried. Total phenol and total flavonoid amounts were investigated; contents of amygdalin (Prunus persica), paeoniflorin (Paeonia lactiflora), and glycyrrhizin (Glycyrrhiza uralensis) - the marker compounds of Danggwisu-san (Dangguixu-san) - were also analyzed through high performance liquid chromatography (HPLC). Results No significant change in total phenol and total flavonoid amounts was observed under the indicated storage conditions. Moreover, the contents of marker compounds, i.e. amygdalin, paeoniflorin, and glycyrrhizin, did not alter significantly under the indicated conditions, as well. Conclusions Danggwisu-san (Dangguixu-san) was found to be stable up until 4 weeks under the indicated conditions. Further studies on efficacy and long-term stability are warranted to establish the expiration period of Danggwisu-san (Dangguixu-san).

Antioxidant and Lipid Peroxidation Inhibition Effects of Catechin Compounds Isolated from Ethyl Acetate Fraction of Grape Seed Ethanol Extract (포도종자 에텔아세테이트 분획물로부터 분리한 카테킨 화합물의 항산화 및 지질과산화 억제효과)

  • Kim, Nan-Young;Park, Sung-Jin;Oh, Deog-Hwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.10
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    • pp.1498-1502
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    • 2005
  • This study evaluated the isolation and identification of biologically active compounds from the ethyl acetate fraction of grape seed extract (Campbell early). Ethyl acetate fraction was further purified with sephadex LP-20 column chromatography. Each biologically active compound for free radical scavenging effect and lipid peroxidation inhibition was isolated and identified with ${1}^H$ and${12}^C$-NMR. Major compounds were identified as (+)-catechin and (-)-epicatechin respectively. The amounts of (+)-catechin and (-)-epicatechin in grape seed were 45.7$\%$ and 35.1$\%$, respectively. The purified (+)-catechin and (-)-epicatechin showed more strong free radical scavenging effects ($RC_{50}$= 11.1 $\mu$g/mL and 10.4 $\mu$g/mL) than ethyl acetate fraction ($RC_{50}$= 15.4 $\mu$g/mL). However, ethyl acetate fraction showed much stronger lipid oxidative inhibition effect than the purified (+)-catechin and (-)-epicatechin.

The Relationship Between Hippuric Acid in Blood Plasma and Toluene Concentration in the Air of Workplace (톨루엔 폭로근로자의 혈장중 마뇨산과 공기중 톨루엔과의 상관관계)

  • Hwang, Cheon-Hyun;Lee, Won-Jin;Chang, Soung-Hoon;Kim, Hyoung-Ah
    • Journal of Preventive Medicine and Public Health
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    • v.33 no.1
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    • pp.45-50
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    • 2000
  • Objectives : This study was undertaken to evaluate correlation between the levels of hippuric acid in blood plasma (HAP) and those of toluene concentration in the workplace air. Methods : Study subjects were composed of two groups; 21 workers who were occupationally exposed to toluene and 25 rural-area residents who were not exposed to any known occupational toluene source, as an exposed group and a reference group, respectively. Mean age and work duration of the exposed was 42 years and five years, respectively. Mean age of the reference was 42 years. To determine toluene concentrations in the workplace air, air sampling has been conducted for more than six hours using a personal sampler, and analyzed by a gas chromatography-flame ionization detector. Concentrations of hippuric acid in biological samples were determined by a high performance liquid chromatography-ultraviolet detector. Results : Geometric mean(geometric standard deviation) of HAP and hippuric acid in urine(HAU) for the exposed was 1.39(2.21) mg/L and 2.77(1.46) g/L, respectively, which were significantly different from those of the reference [HAP, 9.45(2.94); HAU, 0.37(0.45)]. Teluene concentration in the workplace air was 86.92(range: $45.18\sim151.23$)ppm. The level of HAP or HAU was significantly correlated (r=0.70 and r=0.63, respectively) with that of toluene in the workplace air. The estimated regression equation was logHAP(mg/L)=-3.60+1.93 log(toluene, ppm) or logHAU(g/L)=-0.85+0.67 log(toluene, ppm). The magnitude of correlation was further enhanced when analyzing relationship between toluene concentrations lower than 100 ppm and its corresponding HAP levels. Conclusion : Overall, plasma hippuric acid levels were well correlated with toluene concentrations in the workplace air, and a statistically significant correlation was observed for the samples with toluene concentration lower than 100 ppm.

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Simultaneous Determination of Quinolones in Flatfish and Egg Using liquid Chromatography with Fluorescence Detection (액체크로마토그래피를 이용한 광어 및 계란 중 퀴놀론계의 동시분석법 개발)

  • Lee, Sang-Hee;Shim, You-Shin;Kim, Hyun-Ju;Choi, Yoon-Hee;Shin, Dong-Bin
    • Journal of Food Hygiene and Safety
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    • v.23 no.4
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    • pp.324-329
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    • 2008
  • An analytical method for the simultaneous determination of nine quinolones (QNs) namely, marbofloxacin, norfloxacin(IS), ciprofloxacin, danofloxacin, enrofloxacin, sarafloxacin, difloxacin, oxolinic acid, and flumequine in flatfish and egg was developed and validated using liquid chromatography with fluorescence detection (LC-FD). The samples were extracted using a traditional liquid-liquid extraction process; deproteinization was accomplished by the addition of trichloroacetic acid and acetonitrile (ACN), and defatting was performed with hexane. Chromatographic separation was achieved on a reverse phase C8 column with gradient elution using a mobile phase of 200 mM ammonium acetate buffer (pH 4.5) and ACN. The proposed method was validated according to the CODEX guideline. Mean recoveries of QNs from flatfish and egg were 89.6-106.5% with relative standard deviations (RSDs) below 15% at three different concentrations of 50, 100 and $500{\mu}g/kg$. Linearity was obtained with a correlation coefficient ($r^2$) of 0.9989-1.0000. The LOD for the investigated QNs was $1-16{\mu}g/kg$ depending on flatfish and egg. The present method can be applied simultaneously to determine QNs in muscle of flatfish and egg.